KNI-272, a highly selective and potent peptidic HIV protease inhibitor

Kynostatin {KNI-272; systematic name: 3-[3-benzyl-2-hydroxy-9-(isoquinolin-5-yl­oxy)-6-methyl­sulfanyl­methyl-5,8-dioxo-4,7-di­aza­nonanoyl]-N-tert-butyl-1,3-thia­zolane-4-carbox­amide}, a highly selective and potent HIV protease inhibitor containing allo­phenyl­norstatin [(2S,3S)-3-amino-2-hydroxy-4-phenyl­butyric acid], has been crystallized as the hydrate, C₃₃H₄₁N₅O₆S₂·0.803H₂O, from aqueous hexyl­ene glycol. The observed disorder of the phenyl group in the structure is related to the mode of hydration. The backbone conformation of the mol­ecule is twisted and the overall conformation of the free inhibitor is similar to that observed in its complex with HIV protease.

S1. Comment

Kynostatin (KNI-272), hereinafter (I), a peptide mimic containing an allophenylnorstatin residue [Apns, (2S,3S)-3-amino-2-hydroxy-4-phenylbutyric acid; Mimoto et al., 1991], was designed based on the substrate transition state concept for aspartic protease inhibition (Kiso, 1996; Kiso et al., 1999). Compound (I) showed high selectivity and potent inhibitory activity (Ki = 0.0055 nM) for HIV protease (Mimoto et al., 1992), and also displayed good pharmacokinetics and an excellent therapeutic index (Kageyama et al., 1993). Compound (I) is composed of five modules, namely 5-isoquinolyloxy acetic acid (iQoa), methylthioalanine (Mta), Apns, thioproline (Thz) and tert-butyl amine, as shown the Scheme. Crystals of (I) were grown from various alcohol solutions, and the crystal structure obtained from aqueous hexylene glycol is reported here. \sch

The molecular structure of (I) is shown in Fig.1. The phenyl group of the Apns residue is disordered, with two alternate orientations. Analysis of the electron-density map (Fig. 2) shows that rotation of the phenyl ring about the C18—C19 and C19—C20 bonds creates space for the binding of three disordered water molecules whose total site occupancy is 0.8. The water molecules are linked to each other by hydrogen bonds [Table 1; O61···O63 = 3.05 (3) and O63···O62 = 2.67 (3) Å] and interact with the peptide [O62···O32 = 2.77 (2), O61···N1 = 2.932 (12) and O62···N1 = 2.88 (2) Å]. Hydrogen bonds are also formed between the peptides; N18···O27 = 3.038 (6), N33···O26 = 2.984 (7) and O26···O12 = 2.837 (6) Å.

The peptide backbone of (I) is twisted (Fig. 2), and the conformation is different from a β-turn or extended (β-sheet) structure. The structural features of (I) seem to be similar to those of the molecule in the complex with HIV protease (Baldwin et al., 1995). Selected torsion angles are listed in Table 2, to compare the backbone structures of the two compounds. The angles involving the atoms of the Mta residue are significantly different in the free and complex structures (e.g. N13—C13—C14—S15), and the thiazolidine ring is puckered differently (N28—C28—C29—S30). In contrast, the backbone conformation of the Apns-Thz-Tbu moiety observed in the crystal of (I) is closely related to that observed in the enzyme complex. Furthermore, it is known that the range of observed solution conformations of this fragment is rather limited (Ohno et al., 1996). Thus, it is suggested that the Apns-Thz-^tBu fragment has a relatively rigid conformation, which is nearly optimal for tight interactions between the Apns moiety of (I) and the catalytic centre of the HIV protease active site.

Related literature

For related literature, see: Baldwin et al. (1995); Kageyama et al. (1993); Kiso (1996); Kiso et al. (1999); Leslie (1990); Mimoto et al. (1991, 1992); Ohno et al. (1996).

S2. Experimental

Compound (I) was synthesized according to the method published by Mimoto et al. (1991). Compound (I) (20 mg) was dissolved in hexylene glycol (0.3 ml), and water was added to this solution in several small portions of ?ml. The addition of water was stopped before the solution became opaque. The solution was then sealed in a vial and crystals were grown for 2–5 d at room temperature. A crystal of (I) was mounted on a nylon loop with mother liquor solution (aqueous hexylene glycol) and frozen in a nitrogen stream at 100 K.

S3. Refinement

Intensity data were collected by the oscillation method using synchrotron radiation (SPring-8/BL24XU—A). A total of 60 images covered 180° rotation on the ϕ axis. Image data were processed using MOSFLM (Leslie, 1990) (Query year - 1999 given in tables above) and the CCP4 program suite (Collaborative Computational Project, Number 4, 1994). The high background regions were observed at approximately 3.0 Å resolution and these regions were excluded from the processing. A total of 15444 reflections were observed from the image data, and these were averaged to 6157 reflections by the CCP4 program suite, with R_merge = 0.092. The H atoms of the peptide were placed in calculated positions and constrained during the refinement, with C—H = ?Å and U_iso(H) = ?U_eq(C). Please provide details of constraints used. Please also provide CIF-format details of the bond distances and angles involving H atoms. The H atoms of the water molecules were positioned by considering the hydrogen-bonding networks and were fixed during the refinement. Constraints? The absolute stereochemistry is known from the known chirality of the starting materials in the synthesis, but it could not be determined from the present data.

Computing details

Data collection: PROCESS (Rigaku, 1996); cell refinement: MOSFLM (Leslie, 1999); data reduction: MOSFLM; program(s) used to solve structure: SHELXS97 (Sheldrick, 1997); program(s) used to refine structure: SHELXL97 (Sheldrick, 1997); molecular graphics: PLATON (Spek, 1998); software used to prepare material for publication: PARST (Nardelli, 1983).

Figures

	Fig. 1. The molecular view of (I) with displacement ellipsoids drawn at the 30% probability level. Thin and dotted lines show the two disordered structures of the Apns residue and the water molecules.
	Fig. 2. The electron-density map of (I) (Swiss-PdbViewer; Guex & Peitsch, 1997), showing the disordered state of (I) and the water molecules. Contours show the boundaries of 1.0 and 2.5 σ. Three water molecules (O61, O62 and O63) bind when the phenyl ring swings into an alternate conformation designated by the primed atoms.

3-[3-benzyl-2-hydroxy-9-(isoquinolin-5-yloxy)-6-methylsulfanylmethyl-5,8-dioxo- 4,7-diazanonanoyl]-N-tert-butyl-1,3-thiazolane-4-carboxamide

Crystal data

C33H41N5O6S2·0.803H2O	Z = 2
Mr = 682.33	F(000) = 724
Monoclinic, P21	Dx = 1.281 Mg m^−3
a = 10.7631 (4) Å	Synchrotron radiation, λ = 0.83600 Å
b = 13.1751 (4) Å	µ = 0.20 mm^−1
c = 12.5623 (5) Å	T = 100 K
β = 96.887 (2)°	Plate, colourless
V = 1768.54 (11) Å^3	0.25 × 0.10 × 0.02 mm

Data collection

Rigaku RAXIS-IV image-plate detector diffractometer	3321 reflections with I > 2σ(I)
Radiation source: double monochromated beam	Rint = 0.070
Diamond monochromator	θmax = 31.5°, θmin = 8.1°
Detector resolution: 10 pixels mm^-1	h = −13→13
oscillation scans	k = −16→16
5965 measured reflections	l = 0→15
3571 independent reflections

Refinement

Refinement on F^2	Secondary atom site location: difference Fourier map
Least-squares matrix: full	Hydrogen site location: inferred from neighbouring sites
R[F^2 > 2σ(F^2)] = 0.067	H-atom parameters constrained
wR(F^2) = 0.191	w = 1/[σ^2(Fo^2) + (0.1047P)^2 + 2.2778P] where P = (Fo^2 + 2Fc^2)/3
S = 0.99	(Δ/σ)max = 0.020
3571 reflections	Δρmax = 0.81 e Å^−3
504 parameters	Δρmin = −0.61 e Å^−3
1 restraint	Extinction correction: SHELXL97 (Sheldrick, 1997), Fc^*=kFc[1+0.001xFc^2λ^3/sin(2θ)]^-1/4
Primary atom site location: structure-invariant direct methods	Extinction coefficient: 0.30 (3)

Special details

Geometry. All e.s.d.'s (except the e.s.d. in the dihedral angle between two l.s. planes) are estimated using the full covariance matrix. The cell e.s.d.'s are taken into account individually in the estimation of e.s.d.'s in distances, angles and torsion angles; correlations between e.s.d.'s in cell parameters are only used when they are defined by crystal symmetry. An approximate (isotropic) treatment of cell e.s.d.'s is used for estimating e.s.d.'s involving l.s. planes.

Refinement. Refinement of F² against ALL reflections. The weighted R-factor wR and goodness of fit S are based on F², conventional R-factors R are based on F, with F set to zero for negative F². The threshold expression of F² > σ(F²) is used only for calculating R-factors(gt) etc. and is not relevant to the choice of reflections for refinement. R-factors based on F² are statistically about twice as large as those based on F, and R- factors based on ALL data will be even larger.

Fractional atomic coordinates and isotropic or equivalent isotropic displacement parameters (Ų)

	x	y	z	Uiso*/Ueq	Occ. (<1)
N1	0.7262 (5)	−0.3745 (4)	0.4204 (4)	0.0433 (12)
C2	0.6760 (5)	−0.3165 (5)	0.4949 (5)	0.0427 (13)
H2	0.7225	−0.2619	0.5250	0.051*
C3	0.5610 (5)	−0.3344 (4)	0.5277 (4)	0.0341 (11)
H3	0.5323	−0.2937	0.5801	0.041*
C4	0.3666 (5)	−0.4408 (4)	0.5086 (4)	0.0298 (11)
C5	0.2969 (5)	−0.5156 (5)	0.4545 (5)	0.0364 (12)
H5	0.2179	−0.5313	0.4728	0.044*
C6	0.3453 (5)	−0.5698 (5)	0.3699 (4)	0.0368 (12)
H6	0.2963	−0.6189	0.3318	0.044*
C7	0.4615 (5)	−0.5502 (4)	0.3448 (4)	0.0338 (11)
H7	0.4936	−0.5881	0.2919	0.041*
C8	0.6561 (5)	−0.4491 (4)	0.3766 (5)	0.0360 (12)
H8	0.6900	−0.4897	0.3269	0.043*
C9	0.4863 (4)	−0.4148 (4)	0.4815 (4)	0.0275 (10)
C10	0.5346 (5)	−0.4723 (4)	0.3988 (4)	0.0300 (10)
O11	0.3287 (3)	−0.3871 (3)	0.5943 (3)	0.0337 (8)
C11	0.2272 (5)	−0.4309 (4)	0.6415 (4)	0.0364 (12)
H11	0.2379	−0.5040	0.6456	0.044*
H12	0.1494	−0.4169	0.5965	0.044*
C12	0.2195 (5)	−0.3894 (4)	0.7526 (4)	0.0316 (11)
O12	0.1507 (4)	−0.4322 (3)	0.8100 (3)	0.0391 (10)
N13	0.2886 (4)	−0.3067 (3)	0.7833 (3)	0.0296 (9)
H13	0.3263	−0.2753	0.7364	0.041*
C13	0.3017 (5)	−0.2686 (4)	0.8933 (4)	0.0289 (10)
H14	0.2467	−0.3070	0.9354	0.040*
C14	0.4376 (5)	−0.2791 (5)	0.9445 (5)	0.0365 (12)
H15	0.4438	−0.2547	1.0178	0.044*
H16	0.4901	−0.2361	0.9059	0.044*
S15	0.49746 (15)	−0.40805 (12)	0.94521 (14)	0.0483 (5)
C16	0.4082 (8)	−0.4681 (6)	1.0394 (6)	0.0612 (19)
H17	0.4322	−0.5381	1.0474	0.067*
H18	0.3207	−0.4637	1.0137	0.067*
H19	0.4239	−0.4345	1.1075	0.067*
C17	0.2626 (5)	−0.1574 (4)	0.8884 (4)	0.0317 (11)
O17	0.3245 (4)	−0.0945 (3)	0.8440 (4)	0.0428 (10)
N18	0.1570 (4)	−0.1348 (3)	0.9310 (4)	0.0308 (9)
H20	0.1275	−0.1771	0.9737	0.037*
C18	0.0913 (5)	−0.0378 (4)	0.9046 (5)	0.0339 (11)
H21	0.1372	−0.0083	0.8494	0.041*
C19	−0.0320 (13)	−0.0643 (11)	0.8453 (12)	0.035 (3)	0.491 (12)
H22	−0.0793	−0.0023	0.8296	0.042*	0.491 (12)
H23	−0.0778	−0.1056	0.8913	0.042*	0.491 (12)
C20	−0.0243 (13)	−0.1215 (15)	0.7403 (12)	0.037 (3)	0.491 (12)
C21	−0.0496 (17)	−0.224 (2)	0.7378 (16)	0.042 (4)	0.491 (12)
H24	−0.0763	−0.2566	0.7964	0.051*	0.491 (12)
C22	−0.0330 (15)	−0.2769 (14)	0.6416 (15)	0.060 (5)	0.491 (12)
H25	−0.0512	−0.3459	0.6365	0.072*	0.491 (12)
C23	0.0096 (12)	−0.2280 (16)	0.5562 (11)	0.058 (5)	0.491 (12)
H26	0.0230	−0.2645	0.4953	0.070*	0.491 (12)
C24	0.0316 (12)	−0.1286 (15)	0.5605 (11)	0.054 (4)	0.491 (12)
H27	0.0581	−0.0958	0.5016	0.064*	0.491 (12)
C25	0.0150 (4)	−0.0725 (4)	0.6539 (4)	0.047 (3)	0.491 (12)
H28	0.0307	−0.0031	0.6566	0.056*	0.491 (12)
C26	0.1111 (4)	0.0390 (4)	0.9963 (4)	0.0304 (11)
H29	0.2013	0.0475	1.0166	0.036*
O26	0.0571 (4)	0.0052 (4)	1.0870 (4)	0.0531 (14)
H30	−0.0005	0.0345	1.0977	0.053*
C27	0.0557 (5)	0.1417 (4)	0.9576 (4)	0.0285 (10)
O27	−0.0411 (3)	0.1727 (3)	0.9914 (3)	0.0361 (9)
N28	0.1099 (4)	0.1918 (3)	0.8825 (4)	0.0281 (9)
C28	0.0501 (4)	0.2847 (3)	0.8367 (4)	0.0280 (10)
H31	0.0386	0.3329	0.8941	0.039*
C29	0.1391 (5)	0.3298 (4)	0.7624 (5)	0.0393 (13)
H32	0.1297	0.4029	0.7574	0.047*
H33	0.1230	0.3010	0.6910	0.047*
S30	0.29496 (13)	0.29578 (11)	0.82504 (14)	0.0430 (5)
C31	0.2325 (5)	0.1706 (4)	0.8481 (5)	0.0357 (12)
H34	0.2242	0.1305	0.7828	0.043*
H35	0.2859	0.1347	0.9034	0.043*
C32	−0.0768 (5)	0.2602 (4)	0.7725 (4)	0.0314 (11)
O32	−0.0943 (4)	0.1804 (3)	0.7221 (3)	0.0372 (9)
N33	−0.1618 (4)	0.3350 (3)	0.7733 (4)	0.0329 (10)
H36	−0.1396	0.3888	0.8094	0.039*
C33	−0.2910 (6)	0.3319 (5)	0.7165 (6)	0.0452 (14)
C34	−0.3595 (6)	0.2375 (6)	0.7460 (6)	0.0542 (18)
H37	−0.3618	0.2359	0.8222	0.054*
H38	−0.3168	0.1784	0.7245	0.054*
H39	−0.4434	0.2384	0.7101	0.054*
C35	−0.3563 (8)	0.4253 (8)	0.7562 (12)	0.105 (5)
H41	−0.3574	0.4207	0.8323	0.105*
H42	−0.4406	0.4286	0.7213	0.105*
H43	−0.3119	0.4854	0.7397	0.105*
C36	−0.2866 (10)	0.3375 (7)	0.5954 (7)	0.070 (2)
H44	−0.2453	0.2783	0.5722	0.070*
H45	−0.2413	0.3971	0.5788	0.070*
H46	−0.3703	0.3406	0.5592	0.070*
C19'	−0.0615 (12)	−0.0508 (8)	0.8923 (13)	0.033 (3)	0.509 (12)
H23'	−0.0891	−0.0528	0.9630	0.040*	0.509 (12)
H22'	−0.1002	0.0075	0.8545	0.040*	0.509 (12)
C20'	−0.1036 (10)	−0.1475 (8)	0.8314 (10)	0.039 (3)	0.509 (12)
C25'	−0.0433 (18)	−0.176 (2)	0.7441 (18)	0.048 (5)	0.509 (12)
H28'	0.0176	−0.1354	0.7185	0.058*	0.509 (12)
C24'	−0.0816 (18)	−0.271 (2)	0.696 (2)	0.067 (5)	0.509 (12)
H27'	−0.0432	−0.2952	0.6389	0.081*	0.509 (12)
C23'	−0.1733 (15)	−0.3274 (10)	0.7337 (15)	0.066 (5)	0.509 (12)
H26'	−0.1931	−0.3899	0.7016	0.053*	0.509 (12)
C22'	−0.2373 (15)	−0.2978 (12)	0.8147 (10)	0.053 (4)	0.509 (12)
H25'	−0.3032	−0.3357	0.8353	0.064*	0.509 (12)
C21'	−0.1993 (12)	−0.2081 (10)	0.8649 (10)	0.047 (3)	0.509 (12)
H24'	−0.2384	−0.1868	0.9232	0.057*	0.509 (12)
O61	−0.0038 (10)	−0.3377 (13)	0.4640 (9)	0.053 (4)	0.366 (9)
H62	−0.0110	−0.2750	0.4653	0.058*	0.366 (9)
H61	−0.0802	−0.3622	0.4434	0.058*	0.366 (9)
O62	0.0318 (18)	0.043 (2)	0.6072 (17)	0.051 (6)	0.204 (5)
H63	0.1117	0.0352	0.6070	0.056*	0.204 (5)
H64	−0.0014	0.0814	0.6527	0.056*	0.204 (5)
O63	−0.0373 (14)	−0.1078 (14)	0.4713 (13)	0.034 (4)	0.233 (6)
H65	−0.0717	−0.1637	0.4883	0.038*	0.233 (6)
H66	−0.0157	−0.0658	0.5121	0.038*	0.233 (6)

Atomic displacement parameters (Ų)

	U^11	U^22	U^33	U^12	U^13	U^23
N1	0.033 (2)	0.039 (3)	0.061 (3)	−0.010 (2)	0.020 (2)	−0.014 (2)
C2	0.033 (3)	0.037 (3)	0.060 (3)	−0.010 (3)	0.013 (2)	−0.016 (3)
C3	0.030 (2)	0.031 (3)	0.043 (3)	−0.002 (2)	0.013 (2)	−0.005 (2)
C4	0.034 (2)	0.026 (3)	0.032 (2)	−0.001 (2)	0.0124 (19)	−0.001 (2)
C5	0.031 (2)	0.036 (3)	0.044 (3)	−0.011 (2)	0.013 (2)	−0.005 (2)
C6	0.040 (3)	0.033 (3)	0.037 (3)	−0.010 (2)	0.008 (2)	−0.010 (2)
C7	0.038 (3)	0.031 (3)	0.034 (2)	−0.001 (2)	0.009 (2)	−0.002 (2)
C8	0.034 (3)	0.031 (3)	0.046 (3)	0.001 (2)	0.016 (2)	0.000 (2)
C9	0.026 (2)	0.023 (2)	0.033 (2)	−0.001 (2)	0.0051 (18)	0.000 (2)
C10	0.034 (2)	0.022 (2)	0.035 (2)	−0.001 (2)	0.0077 (19)	0.000 (2)
O11	0.0347 (18)	0.0295 (19)	0.0401 (18)	−0.0083 (16)	0.0174 (15)	−0.0056 (16)
C11	0.034 (2)	0.034 (3)	0.045 (3)	−0.013 (2)	0.022 (2)	−0.011 (2)
C12	0.031 (2)	0.026 (2)	0.042 (3)	−0.004 (2)	0.018 (2)	−0.004 (2)
O12	0.042 (2)	0.0281 (19)	0.053 (2)	−0.0109 (16)	0.0280 (17)	−0.0083 (17)
N13	0.034 (2)	0.027 (2)	0.0289 (19)	−0.0050 (18)	0.0088 (16)	0.0026 (17)
C13	0.033 (2)	0.022 (2)	0.034 (2)	−0.002 (2)	0.0125 (19)	0.0003 (19)
C14	0.033 (3)	0.034 (3)	0.042 (3)	0.001 (2)	0.003 (2)	0.001 (2)
S15	0.0432 (8)	0.0378 (8)	0.0632 (9)	0.0111 (7)	0.0034 (7)	−0.0015 (7)
C16	0.079 (5)	0.043 (4)	0.060 (4)	0.006 (4)	0.005 (4)	0.013 (3)
C17	0.037 (3)	0.023 (2)	0.035 (2)	−0.005 (2)	0.007 (2)	0.002 (2)
O17	0.053 (2)	0.0247 (19)	0.052 (2)	−0.0087 (18)	0.0136 (19)	0.0083 (18)
N18	0.037 (2)	0.0164 (18)	0.040 (2)	0.0005 (17)	0.0062 (17)	0.0039 (17)
C18	0.036 (3)	0.020 (2)	0.045 (3)	−0.002 (2)	−0.002 (2)	0.003 (2)
C19	0.031 (6)	0.030 (6)	0.040 (7)	0.003 (5)	−0.008 (5)	−0.014 (6)
C20	0.023 (6)	0.036 (8)	0.050 (7)	0.005 (6)	0.003 (4)	−0.003 (7)
C21	0.030 (7)	0.046 (12)	0.048 (8)	−0.006 (11)	−0.004 (6)	−0.015 (11)
C22	0.038 (7)	0.066 (10)	0.074 (10)	0.005 (7)	−0.002 (7)	−0.035 (9)
C23	0.037 (6)	0.093 (13)	0.043 (7)	0.016 (7)	−0.003 (5)	−0.030 (8)
C24	0.031 (6)	0.088 (12)	0.042 (6)	0.007 (7)	0.006 (5)	−0.019 (7)
C25	0.034 (6)	0.068 (9)	0.038 (6)	0.011 (6)	0.004 (4)	−0.004 (6)
C26	0.028 (2)	0.023 (2)	0.042 (3)	0.004 (2)	0.017 (2)	0.007 (2)
O26	0.061 (3)	0.046 (3)	0.061 (3)	0.034 (2)	0.044 (2)	0.032 (2)
C27	0.030 (2)	0.023 (2)	0.035 (2)	0.0024 (19)	0.0119 (18)	0.000 (2)
O27	0.0364 (19)	0.0289 (19)	0.047 (2)	0.0151 (16)	0.0217 (17)	0.0110 (17)
N28	0.0277 (19)	0.0175 (19)	0.042 (2)	0.0017 (16)	0.0149 (17)	0.0018 (17)
C28	0.030 (2)	0.014 (2)	0.040 (2)	0.0020 (18)	0.0077 (19)	−0.0002 (19)
C29	0.043 (3)	0.024 (2)	0.053 (3)	−0.003 (2)	0.015 (2)	0.006 (2)
S30	0.0321 (7)	0.0322 (8)	0.0660 (10)	−0.0076 (5)	0.0113 (6)	0.0108 (7)
C31	0.029 (2)	0.028 (3)	0.053 (3)	0.000 (2)	0.017 (2)	0.006 (2)
C32	0.035 (3)	0.020 (2)	0.041 (3)	0.002 (2)	0.012 (2)	0.004 (2)
O32	0.0364 (19)	0.0243 (18)	0.052 (2)	−0.0006 (16)	0.0112 (17)	−0.0104 (17)
N33	0.036 (2)	0.021 (2)	0.041 (2)	0.0022 (18)	0.0002 (18)	−0.0033 (18)
C33	0.039 (3)	0.032 (3)	0.061 (4)	0.007 (2)	−0.010 (3)	−0.014 (3)
C34	0.040 (3)	0.065 (5)	0.061 (4)	−0.011 (3)	0.018 (3)	−0.023 (4)
C35	0.051 (4)	0.072 (6)	0.178 (11)	0.024 (4)	−0.043 (6)	−0.078 (7)
C36	0.090 (6)	0.047 (4)	0.065 (4)	−0.014 (4)	−0.030 (4)	0.019 (4)
C19'	0.037 (6)	0.012 (5)	0.049 (8)	0.005 (4)	−0.005 (5)	0.001 (5)
C20'	0.035 (5)	0.024 (5)	0.057 (7)	0.008 (4)	−0.001 (5)	0.009 (5)
C25'	0.029 (8)	0.033 (11)	0.080 (12)	0.001 (11)	−0.007 (7)	−0.002 (13)
C24'	0.043 (9)	0.084 (15)	0.072 (12)	0.034 (11)	−0.004 (9)	−0.004 (12)
C23'	0.060 (9)	0.022 (6)	0.106 (12)	0.002 (6)	−0.033 (9)	0.013 (7)
C22'	0.064 (8)	0.049 (8)	0.042 (6)	−0.002 (7)	−0.010 (6)	0.005 (6)
C21'	0.052 (7)	0.034 (6)	0.050 (6)	−0.007 (6)	−0.016 (5)	0.011 (6)
O61	0.026 (5)	0.091 (11)	0.045 (6)	0.004 (6)	0.015 (5)	0.007 (7)
O62	0.025 (9)	0.084 (18)	0.048 (11)	0.003 (11)	0.017 (8)	0.003 (12)
O63	0.027 (7)	0.043 (10)	0.035 (8)	0.001 (7)	0.011 (6)	−0.015 (7)

Geometric parameters (Å, º)

N1—C8	1.318 (8)	O61—O62^i	1.82 (3)
N1—C2	1.369 (8)	O62—O61^ii	1.82 (3)
C2—C3	1.371 (7)	C2—H2	0.930 (6)
C3—C9	1.412 (7)	C3—H3	0.930 (5)
C4—C5	1.369 (8)	C5—H5	0.930 (6)
C4—O11	1.389 (6)	C6—H6	0.930 (6)
C4—C9	1.414 (7)	C7—H7	0.930 (5)
C5—C6	1.429 (8)	C8—H8	0.929 (6)
C6—C7	1.351 (7)	C11—H11	0.971 (5)
C7—C10	1.416 (8)	C11—H12	0.970 (5)
C8—C10	1.403 (7)	N13—H13	0.860 (4)
C9—C10	1.432 (7)	C13—H14	0.980 (6)
O11—C11	1.426 (6)	C14—H15	0.970 (6)
C11—C12	1.511 (7)	C14—H16	0.970 (6)
C12—O12	1.230 (6)	C16—H17	0.960 (8)
C12—N13	1.348 (7)	C16—H18	0.960 (8)
N13—C13	1.462 (7)	C16—H19	0.960 (8)
C13—C17	1.524 (7)	N18—H20	0.860 (5)
C13—C14	1.531 (7)	C18—H21	0.979 (6)
C14—S15	1.817 (6)	C19—H22	0.970 (14)
S15—C16	1.794 (8)	C19—H23	0.970 (15)
C17—O17	1.238 (7)	C21—H24	0.93 (2)
C17—N18	1.347 (7)	C22—H25	0.930 (18)
N18—C18	1.478 (7)	C23—H26	0.929 (16)
C18—C19	1.484 (13)	C24—H27	0.931 (16)
C18—C26	1.529 (7)	C25—H28	0.930 (5)
C18—C19'	1.642 (14)	C26—H29	0.980 (4)
C19—C20	1.53 (2)	O26—H30	0.756 (5)
C20—C25	1.372 (17)	C28—H31	0.980 (5)
C20—C21	1.37 (3)	C29—H32	0.970 (5)
C21—C22	1.43 (3)	C29—H33	0.970 (6)
C22—C23	1.38 (3)	C31—H34	0.971 (6)
C23—C24	1.33 (3)	C31—H35	0.970 (6)
C24—C25	1.416 (14)	N33—H36	0.860 (4)
C26—O26	1.412 (6)	C34—H37	0.961 (8)
C26—C27	1.535 (7)	C34—H38	0.959 (8)
C27—O27	1.240 (6)	C34—H39	0.960 (6)
C27—N28	1.341 (6)	C35—H41	0.959 (15)
N28—C31	1.465 (6)	C35—H42	0.960 (9)
N28—C28	1.468 (6)	C35—H43	0.960 (11)
C28—C32	1.535 (7)	C36—H44	0.960 (10)
C28—C29	1.535 (7)	C36—H45	0.960 (10)
C29—S30	1.822 (6)	C36—H46	0.960 (10)
S30—C31	1.817 (6)	C19'—H23'	0.970 (17)
C32—O32	1.230 (7)	C19'—H22'	0.970 (12)
C32—N33	1.346 (7)	C25'—H28'	0.93 (2)
N33—C33	1.485 (7)	C24'—H27'	0.93 (3)
C33—C34	1.514 (10)	C23'—H26'	0.930 (14)
C33—C36	1.530 (12)	C22'—H25'	0.929 (16)
C33—C35	1.531 (10)	C21'—H24'	0.931 (13)
C19'—C20'	1.528 (18)	O61—H62	0.830 (17)
C20'—C25'	1.39 (3)	O61—H61	0.892 (12)
C20'—C21'	1.406 (18)	O62—H63	0.87 (2)
C25'—C24'	1.43 (3)	O62—H64	0.87 (2)
C24'—C23'	1.36 (3)	O63—H65	0.863 (18)
C23'—C22'	1.35 (2)	O63—H66	0.771 (17)
C22'—C21'	1.38 (2)
C8—N1—C2	116.7 (5)	O11—C11—H12	109.4 (5)
N1—C2—C3	123.6 (5)	C12—C11—H11	109.2 (5)
C2—C3—C9	119.7 (5)	C12—C11—H12	109.3 (5)
C5—C4—O11	123.8 (4)	H11—C11—H12	107.9 (5)
C5—C4—C9	121.0 (4)	C12—N13—H13	118.8 (5)
O11—C4—C9	115.1 (4)	C13—N13—H13	118.8 (4)
C4—C5—C6	120.1 (5)	N13—C13—H14	109.7 (4)
C7—C6—C5	120.6 (5)	C14—C13—H14	109.6 (5)
C6—C7—C10	120.2 (5)	C17—C13—H14	109.7 (5)
N1—C8—C10	125.4 (5)	C13—C14—H15	108.8 (5)
C3—C9—C4	125.0 (5)	C13—C14—H16	108.8 (5)
C3—C9—C10	117.1 (4)	S15—C14—H15	108.7 (5)
C4—C9—C10	117.9 (5)	S15—C14—H16	108.7 (5)
C8—C10—C7	122.7 (5)	H15—C14—H16	107.6 (6)
C8—C10—C9	117.2 (5)	S15—C16—H17	109.5 (6)
C7—C10—C9	120.1 (4)	S15—C16—H18	109.4 (7)
C4—O11—C11	115.3 (4)	S15—C16—H19	109.5 (6)
O11—C11—C12	111.5 (4)	H17—C16—H18	109.5 (8)
O12—C12—N13	123.6 (5)	H17—C16—H19	109.5 (8)
O12—C12—C11	118.5 (5)	H18—C16—H19	109.4 (8)
N13—C12—C11	117.8 (4)	C17—N18—H20	119.8 (5)
C12—N13—C13	122.3 (4)	C18—N18—H20	119.8 (4)
N13—C13—C17	107.3 (4)	N18—C18—H21	103.5 (5)
N13—C13—C14	110.2 (4)	C19—C18—H21	103.6 (7)
C17—C13—C14	110.4 (4)	C26—C18—H21	103.6 (5)
C13—C14—S15	114.0 (4)	C18—C19—H22	108.7 (12)
C16—S15—C14	101.4 (3)	C18—C19—H23	108.7 (12)
O17—C17—N18	123.8 (5)	C20—C19—H22	108.7 (13)
O17—C17—C13	120.1 (5)	C20—C19—H23	108.7 (13)
N18—C17—C13	116.0 (4)	H22—C19—H23	107.6 (14)
C17—N18—C18	120.4 (4)	C20—C21—H24	121 (2)
N18—C18—C19	106.5 (7)	C22—C21—H24	121.9 (19)
N18—C18—C26	112.6 (4)	C21—C22—H25	119.3 (19)
C19—C18—C26	124.7 (8)	C23—C22—H25	119.4 (18)
N18—C18—C19'	112.2 (5)	C22—C23—H26	119.9 (14)
C19—C18—C19'	26.2 (5)	C24—C23—H26	119.9 (18)
C26—C18—C19'	100.8 (7)	C23—C24—H27	119.7 (17)
C18—C19—C20	114.2 (12)	C25—C24—H27	119.8 (12)
C25—C20—C21	121.5 (16)	C20—C25—H28	120.4 (9)
C25—C20—C19	120.3 (15)	C24—C25—H28	120.2 (9)
C21—C20—C19	118.1 (15)	C18—C26—H29	108.4 (4)
C20—C21—C22	117 (2)	O26—C26—H29	108.5 (4)
C23—C22—C21	121.0 (19)	C27—C26—H29	108.5 (4)
C24—C23—C22	120.3 (13)	C26—O26—H30	114.1 (5)
C23—C24—C25	120.5 (14)	N28—C28—H31	109.9 (4)
C20—C25—C24	119.4 (12)	C29—C28—H31	109.9 (4)
O26—C26—C18	111.4 (5)	C32—C28—H31	109.9 (4)
O26—C26—C27	110.6 (4)	C28—C29—H32	110.8 (5)
C18—C26—C27	109.3 (4)	C28—C29—H33	110.8 (5)
O27—C27—N28	122.2 (5)	S30—C29—H32	110.8 (4)
O27—C27—C26	119.5 (4)	S30—C29—H33	110.9 (5)
N28—C27—C26	118.2 (4)	H32—C29—H33	108.8 (5)
C27—N28—C31	126.7 (4)	N28—C31—H34	111.0 (5)
C27—N28—C28	118.7 (4)	N28—C31—H35	111.1 (5)
C31—N28—C28	114.3 (4)	S30—C31—H34	110.9 (4)
N28—C28—C32	110.6 (4)	S30—C31—H35	111.0 (4)
N28—C28—C29	106.6 (4)	H34—C31—H35	109.1 (6)
C32—C28—C29	110.1 (4)	C32—N33—H36	117.5 (4)
C28—C29—S30	104.6 (4)	C33—N33—H36	117.4 (5)
C31—S30—C29	87.3 (3)	C33—C34—H37	109.5 (7)
N28—C31—S30	103.7 (4)	C33—C34—H38	109.5 (7)
O32—C32—N33	124.6 (5)	C33—C34—H39	109.5 (6)
O32—C32—C28	121.4 (5)	H37—C34—H38	109.5 (7)
N33—C32—C28	113.9 (4)	H37—C34—H39	109.5 (7)
C32—N33—C33	125.1 (5)	H38—C34—H39	109.5 (7)
N33—C33—C34	111.1 (6)	C33—C35—H41	109.6 (10)
N33—C33—C36	109.7 (6)	C33—C35—H42	109.5 (8)
C34—C33—C36	110.9 (6)	C33—C35—H43	109.4 (9)
N33—C33—C35	105.2 (5)	H41—C35—H42	109.5 (12)
C34—C33—C35	109.0 (7)	H41—C35—H43	109.5 (12)
C36—C33—C35	110.8 (8)	H42—C35—H43	109.4 (9)
C20'—C19'—C18	111.6 (10)	C33—C36—H44	109.5 (8)
C25'—C20'—C21'	120.9 (16)	C33—C36—H45	109.4 (8)
C25'—C20'—C19'	118.6 (16)	C33—C36—H46	109.5 (8)
C21'—C20'—C19'	120.5 (12)	H44—C36—H45	109.4 (9)
C20'—C25'—C24'	115 (2)	H44—C36—H46	109.4 (10)
C23'—C24'—C25'	121 (2)	H45—C36—H46	109.5 (10)
C22'—C23'—C24'	123.9 (18)	C20'—C19'—H23'	109.3 (12)
C23'—C22'—C21'	116.2 (14)	C20'—C19'—H22'	109.3 (9)
C22'—C21'—C20'	122.4 (14)	H23'—C19'—H22'	108.0 (13)
N1—C2—H2	118.2 (6)	C20'—C25'—H28'	122.1 (19)
C3—C2—H2	118.2 (6)	C24'—C25'—H28'	122 (2)
C2—C3—H3	120.1 (5)	C25'—C24'—H27'	120 (2)
C9—C3—H3	120.2 (5)	C23'—C24'—H27'	119 (2)
C4—C5—H5	120.0 (6)	C24'—C23'—H26'	118.0 (16)
C6—C5—H5	119.9 (6)	C22'—C23'—H26'	118.1 (15)
C5—C6—H6	119.7 (6)	C23'—C22'—H25'	121.9 (15)
C7—C6—H6	119.8 (6)	C21'—C22'—H25'	121.9 (15)
C6—C7—H7	119.8 (5)	C20'—C21'—H24'	118.7 (12)
C10—C7—H7	119.9 (5)	C22'—C21'—H24'	118.8 (13)
N1—C8—H8	117.3 (6)	H62—O61—H61	106.3 (15)
C10—C8—H8	117.3 (5)	H63—O62—H64	124 (2)
O11—C11—H11	109.4 (5)	H65—O63—H66	124 (2)

Symmetry codes: (i) −x, y−1/2, −z+1; (ii) −x, y+1/2, −z+1.

Hydrogen-bond geometry (Å, º)

D—H···A	D—H	H···A	D···A	D—H···A
O26—H30···O27	0.76	2.269	2.669 (6)	114
O62—H64···O32	0.87	1.916	2.77 (2)	166
O61—H62···O63	0.83	2.223	3.05 (3)	178
O63—H66···O62	0.77	1.897	2.67 (3)	177
N18—H20···O27^iii	0.86	2.25	3.038 (6)	152
N33—H36···O26^iv	0.86	2.14	2.984 (7)	169
O26—H30···O12^iv	0.76	2.145	2.837 (6)	153
O61—H61···N1^v	0.89	2.075	2.932 (12)	161
O62—H63···N1^vi	0.87	2.173	2.88 (2)	139

Symmetry codes: (iii) −x, y−1/2, −z+2; (iv) −x, y+1/2, −z+2; (v) x−1, y, z; (vi) −x+1, y+1/2, −z+1.